他扎罗汀对培养角质形成细胞表达RAR和RXR mRNA的影响

目的了解他扎罗汀对对数生长期角质形成细胞表达RAR和RXR mRNA的影响,探讨他扎罗汀治疗银屑病的可能机制。方法用原位杂交法对他扎罗汀干预不同时间段培养的角质形成细胞进行RAR和RXR mRNA染色,然后对各实验组测定数据进行统计学分析。结果他扎罗汀能在24,48,72h三个时间段显著降低RAR和RXR mR-NA的表达。结论他扎罗汀在治疗银屑病过程中可能不仅仅是通过抑制角质形成细胞的增殖达到疗效。     

他扎罗汀的安全性、有效性及其临床应用

在美国他扎罗汀 (ｔａｚａｒｏｔｅｎｅ)是唯一适用于局部治疗斑块状银屑病的维甲酸 ,也是治疗寻常型痤疮的最新局部外用维甲酸。它治疗银屑病的机制是通过调节 3项病理特征 :角质形成细胞的异常分化 ,角质形成细胞的高度增生以及炎症。对寻常型痤疮的作用则是使形成小粉刺的毛囊异常角化上皮恢复正常 ,并且改善毛囊微环境 ,减少痤疮丙酸杆菌的繁殖。1　药物动力学吸收 :他扎罗汀局部应用大多保留在皮肤表面或皮肤内 ,在银屑病皮损处非封包外用 10小时后吸收量不足 1%,而正常皮肤封包 10小时吸收量不足 6 %。代谢 :他扎罗汀是前体药物 ,在皮…     

他扎罗汀诱导前后角质形成细胞TIG 3 mRNA表达

目的研究他扎罗汀对培养的正常人角质形成细胞增殖和他扎罗汀诱导基因3(TIG3)mRNA表达的影响。方法用0.1和1.0μmol/L他扎罗汀处理正常人角质形成细胞24h后,用MTT法、RT-PCR和实时定量RT-PCR(TaqMan探针)法分别检测细胞增殖和TIG3mRNA表达的改变。结果他扎罗汀可抑制角质形成的增殖和诱导TIG3mRNA的表达。0.1和1.0μmol/L他扎罗汀处理角质形成细胞后,分别使角质形成细胞增殖抑制5.0%和11.1%,使TIG3mRNA增加到正常对照组的2.1倍和2.8倍。结论他扎罗汀可抑制角质形成细胞的增殖和上调TIG3mRNA的表达。     

他扎罗汀和NB-UVB对角质形成细胞他扎罗汀诱导基因1 mRNA的影响

目的研究他扎罗汀和窄谱中波紫外线(NB-UVB)对培养的正常人角质形成细胞他扎罗汀诱导基因1(TIG1)mRNA表达的影响。方法用10-7~10-6mol/L他扎罗汀和/或50~100mJ/cm2NB-UVB处理培养的正常人角质形成细胞24h后,用普通RT-PCR和实时荧光定量RT-PCR法检测角质形成细胞TIG1mRNA水平的改变。结果正常人角质形成细胞可表达少量的TIG1mRNA,10-7mol/L和10-6mol/L他扎罗汀单独处理角质形成细胞24h后,TIG1mRNA的水平升高;50mJ/cm2和100mJ/cm2NB-UVB单独照射角质形成细胞24h后,TIG1 mRNA的水平无明显变化;二者联合作用时,TIG1mRNA的水平明显升高,且高于二者单独处理时的水平(P<0.01)。结论他扎罗汀和NB-UVB联合作用时可能通过协同上调TIG1的表达而发挥二者对角质形成细胞功能的调节作用。     

他扎罗汀对Colo-16细胞体外增殖的影响及其机制的研究

目的:研究他扎罗汀对体外培养的皮肤鳞状细胞癌细胞株Colo-16细胞体外增殖的影响,并探讨其机制.方法:用不同浓度的他扎罗汀处理Colo-16细胞,四甲基偶氮唑蓝(MTT)法测定Colo-16细胞增殖活性;免疫组化染色技术及免疫印迹法分别测定Colo-16细胞Bcl-2、Bax蛋白表达水平.结果:不同浓度的他扎罗汀对Colo-16细胞体外增殖均具有抑制作用,其抑制作用呈明显的时效和量效关系(P<0.05);他扎罗汀作用Colo-16细胞后,Bcl-2蛋白表达明显下调,Bax表达明显增强.结论:他扎罗汀能抑制皮肤鳞状细胞癌Colo-16细胞的增殖,其机制与Bcl-2蛋白表达下调和Bax表达增加有关.     

他扎罗汀联合干扰素γ对角质形成细胞HLA-DR表达的影响

目的探讨他扎罗汀对角质形成细胞(KC)增殖及他扎罗汀、干扰素(IFN)-γ及二者联合对HLA-DR在KC上表达的影响,为他扎罗汀的抗增殖作用及其对炎症的影响进行初探。方法①体外培养正常人KC;②用MTT法检测他扎罗汀对KC增殖的影响;③用免疫组化的方法,分别观察他扎罗汀组、IFN-γ组及二者联合组对HLA-DR在KC上表达的影响。结果①10-7~10-5mol/L他扎罗汀作用KC24h、48h后,处理组的细胞增殖较对照组显著降低且呈剂量依赖性;②正常人KC基本不表达HLA-DR;③10-6mol/L他扎罗汀作用24h后,不能诱导KC表达HLA-DR;④500U/mLIFN-γ处理24h后可明显诱导KC表达HLA-DR;⑤10-7~10-5mol/L他扎罗汀联合IFN-γ组作用24h后,KC上HLA-DR的表达诱导作用较单独IFN-γ组显著增强(P<0.005),且呈剂量依赖性。结论他扎罗汀对体外培养的KC增殖有抑制作用;他扎罗汀对IFN-γ诱导KC表达HLA-DR有增强作用。     

外用他扎罗汀凝胶前后PAFR在银屑病患者表皮中的表达

目的了解局部外用0.05%他扎罗汀凝胶前后PAFR在银屑病患者表皮中表达变化,探讨PAFR在他扎罗汀治疗银屑病过程中的可能作用。方法免疫组化法对皮肤组织进行PAFR染色。结果用药前皮损中,13例的PAFR染色为弱阳性,2例为无明显染色,用药1周后,11例皮损的PAFR染色强度明显增强。结论他扎罗汀治疗银屑病可上调皮肤中的PAFR表达,是治疗银屑病的分子生物学机理之一。     

他扎罗汀凝胶治疗寻常型银屑病的临床观察

他扎罗汀是一种新型的受体选择性维A酸类药物犤1犦。其治疗银屑病的主要作用机制是调节角质形成细胞(KC)的分化异常,改善KC的过度增殖,促进炎症消退犤2犦。为进一步验证他扎罗汀凝胶治疗寻常型银屑病的疗效及安全性,笔者用他扎罗汀凝胶(重庆华邦制药有限公司)治疗了115例寻常型银屑病患者,并进行了临床观察,疗效满意,现报告如下。1对象与方法1.1观察对象为2001年10月～2002年1月在上海市12家医院就诊的寻常型银屑病患者,共115例。病人平均年龄45岁(18～65岁),病程6个月～20年。皮损面积占整个体表的2%～30%。靶皮损基线评分:斑块肥厚程度…     

他扎罗汀凝胶治疗寻常性银屑病疗效观察

目的　探讨他扎罗汀治疗寻常性银屑病的临床疗效。方法　两组患者各为 45例 ,分别用他扎罗汀和氯松霜外用 ,根据皮损变化判定疗效。结果　他扎罗汀治疗寻常性银屑病有效率 82 .2 % ,斑块型的有效率 92 .0 % ,均高于氯松霜组 ( 6 2 .2 %、6 0 .7% )。结论　他扎罗汀治疗寻常性银屑病疗效较好 ,尤其适用于斑块型。     

窄谱中波紫外线联合他扎罗汀治疗的寻常性银屑病皮损他扎罗汀诱导基因2的表达

目的 探讨NB-UVB联合他扎罗汀治疗寻常性银屑病的作用机制。方法 用免疫组化和原位杂交的方法检测NB-UVB和（或）他扎罗汀治疗前后银屑病皮损及未受累皮肤中TIG2蛋白和m RNA水平的变化。结果 治疗前，银屑病皮损中几乎无TIG2的表达，单用NB-UVB治疗2周后，角化不全的角质层和棘层上部可见TIG2轻度着色，治疗6周后，可见TIG2阳性表达于棘层上中部。两者联合治疗2周后，可见棘层全层和基底层着色，且在棘层上部着色重于中下部，6周后则表达于表皮全层，且表皮变薄。用NB-UVB和（或）他扎罗汀治疗后，TIG2阳性表达起始于角化不全的角质层和棘层上部至中下部，直至基底层，而他扎罗汀和NB-UVB联合应用后可加速此过程。结论 NB-UVB联合他扎罗汀协同治疗银屑病的机制可能与上调银屑病皮损中TIG2的表达有关。     

Expression of vascular endothelial growth factor in normal epidermis, epithelial tumors and cultured keratinocytes

Vascular endothelial growth factor (VEGF), an endothelium-specific growth factor and microvessel hypermeability factor, is expressed and secreted by several kinds of cells and is implicated in angiogenesis of tumors. The present study was performed to determine the relationship between the expression of VEGF in normal skin, benign and malignant epithelial lesions and cultured keratinocytes and the proliferative activity and degree of differentiation of keratinocytes. Skin lesions were studied immunohistochemically by staining with two anti-VEGF antibodies and secretion and production of VEGF by keratinocyte cultures were evaluated using an enzyme-linked immunosorbent assay. Low to moderate VEGF expression was observed in normal epidermis. In epithelial tumors, different reactivity patterns were observed and different areas of the same tumor expressed different amounts of VEGF. A more prominent labelling occurred in proliferative layers and/or more differentiated cells of virus-induced lesions, squamous cell carcinomas and Bowen’s disease, whereas basal cell carcinomas always stained weakly for VEGF. In cultured keratinocytes, the amount of cell-associated and secreted VEGF increased with time, and the constitutively produced VEGF was mostly released extracellularly. High calcium concentrations upregulated the intracellular content of VEGF but downregulated its release. Taken together, these results showed a modulated expression and release of VEGF in relation to the stage of cell differentiation and in rapidly growing or activated keratinocytes. Received: 22 April 1996     

卡泊三醇、维A酸及雷公藤内酯醇对角质形成细胞及COLO16细胞中血管内皮生长因子生成的影响

目的 探讨卡泊三醇、维A酸及雷公藤内酯醇对角质形成细胞及COLO16细胞中血管内皮生长因子(VEGF)生成的影响.方法 采用RT-PCR法检测原代培养的人角质形成细胞及COLO16细胞中VEGFmRNA的水平.结果 卡泊三醇作用于正常人角质形成细胞4h和24h后,以及作用COLO16细胞24h后,均可抑制VEGFmRNA的表达,呈剂量依赖性,IC₅₀值分别为1.19×10^-4μg/mL、1.51×10^-4μg/mL和3.16×10^-5μg/mL.维A酸作用于正常人角质形成细胞4h后可抑制VEGFmRNA的表达,作用于正常角质形成细胞24h后及作用于COLO16细胞4h和24h均无抑制作用.雷公藤内酯醇对正常角质形成细胞和COLO16细胞中VEGFmRNA表达均无抑制作用.结论 在转录水平抑制VEGF生成可能是维A酸与卡泊三醇抗银屑病的作用机制之一.     

银屑病患者角质形成细胞VEGF表达的研究

目的　研究银屑病发病与血管内皮生长因子 (VEGF)的关系 ,探讨银屑病可能的发病机制。方法　①用免疫组化法检测银屑病患者皮损和非皮损处皮肤、正常健康人皮肤及体外培养的银屑病患者和正常人角质形成细胞(KC)VEGF的表达 ;②用双抗体夹心ELISA法检测银屑病患者及正常人KC培养上清液中VEGF含量。结果　①银屑病皮损处VEGF表达明显高于非皮损处和正常人皮肤 (P均 <0 .0 0 1) ,非皮损处与正常人皮肤VEGF表达也有显著性差异 (P <0 .0 5 ) ;②体外培养的银屑病皮损处和非皮损处KCVEGF表达明显高于正常人 (P均 <0 .0 0 1) ;银屑病皮损处KC与非皮损处KC相比VEGF表达也有显著性差异 (P <0 .0 5 ) )。结论　VEGF可能参与银屑病的发病。     

Vascular endothelial growth factor 121 is the predominant isoform in psoriatic scales

Vascular endothelial growth factor (VEGF) is a powerful agent that causes hyperpermeability of blood vessels as well as endothelial cell proliferation. Recent investigations have revealed that production of VEGF increases in epidermis of psoriatic lesions, and the overproduced VEGF plays an important role in the pathogenesis of psoriasis. In this study, we used immunohistochemical staining as well as extraction of stratum corneum with physiological saline to further analyse VEGF produced in psoriatic lesions. Biological activity of VEGF in the psoriatic scales was assayed by cultured human umbilical vein endothelial cells in vitro. The immunohistochemical examination confirmed an increased production of VEGF in the keratinocytes of psoriatic lesions. In addition, we found that the content of VEGF contained in the psoriatic scales was approximately 50 times greater than that in normal stratum corneum. We also found that VEGF 121 isoform, which has an exclusive ability to cause hyperpermeability of blood vessels, was predominantly detected in psoriatic scales, suggesting a major role of VEGF 121 isoform on the altered structure of microvessels in psoriatic lesions.     

抗角蛋白自身抗体对培养角朊细胞产生白细胞介素8的影响

为探讨抗角蛋白自身抗体（AKantoAb）对角朊细胞免疫功能的影响，实验以IL-1β刺激无血清培养角朊细胞及AKautoAb作用后的角朊细胞，用夹心ELISA法检测培养上清中IL－8的产量。结果表明IL-1β可刺激角朊细胞产生IL－8，并对IL－1β有浓度依赖性，AKautoAb对角朊细胞产生IL－8有明显抑制作用。提示AKautoAb在炎性皮损角朊细胞中的沉着，可能存在有益的调节作用。     

IL-4 and interferon-gamma differentially modulate vascular endothelial growth factor release from normal human keratinocytes and fibroblasts

IL-4 and interferon-gamma (IFN-gamma) are crucial modulators of the immune system and are reported as active antitumor agents and potent inhibitors of angiogenesis. We investigated the effects of these two cytokines on the expression of vascular endothelial growth factor (VEGF), a mediator of major importance in the angiogenesis associated with inflammation, wound healing and tumor invasion and expressed by activated keratinocytes and dermal fibroblasts. Human keratinocytes (HK) and fibroblasts (HF) derived from foreskins, were further cultured during 24 h in defined medium, supplemented or not with the selected growth factors, EGF and TGF-beta1, respectively, before receiving the addition of either IL-4 or IFN-gamma during 24 and 48 h. In basal conditions, fibroblasts produced smaller amounts of VEGF than keratinocytes; the addition of growth factors to the skin cells induced a drastic increase of VEGF secretion. In HF, the basal level of VEGF secretion was reduced by IFN-gamma and slightly increased by IL-4 whereas in HK, IFN-gamma enhanced the secretion of VEGF after 48 h and IL-4 either tended to reduce VEGF secretion or did not exert any effect. Similar but more significant results were observed in skin cells activated by growth-stimulating factors. The association of IL-4 and IFN-gamma mimicked the effects of IFN-gamma alone both in HF and HK. Taken together, these results indicate opposite effects of IFN-gamma and IL-4 on VEGF expression from normal and activated HF and HK. IL-4 may be considered as a poor modulator of VEGF secretion by dermal and epidermal cells. Conversely, IFN-gamma appears as a prominent and versatile mediator in the desregulated angiogenesis associated with inflammatory skin reactions characterized by a T-helper type 1 cell-mediated response.